Rapid detection of Pseudomonas aeruginosa in mouse feces by colorimetric loop-mediated isothermal amplification
Rapid detection of Pseudomonas aeruginosa in mouse feces by colorimetric loop-mediated isothermal amplification
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DOI：
10.1016/j.mimet.2010.03.008
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发表时间：
2010-06-01
影响因子：
2.2
通讯作者：
Hanaki, Ken-Ichi
中科院分区：
文献类型：
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作者：
Goto, Motoki;Shimada, Kayo;Hanaki, Ken-Ichi
建立了羟基萘酚蓝比色环介导等温扩增（LAMP）方法，对铜绿假单胞菌外膜脂蛋白（oprL）基因进行扩增。LAMP检测对血清群和其他细菌的特异性为100%，灵敏度是PCR检测的10倍。LAMP法可以检测到接种于小鼠粪便中的铜绿假单胞菌，接种量为130菌落形成单位(CFU)/0.1 g粪便（3.25 CFU/反应）。DNA提取后2小时内完成检测。在现场试验中，LAMP检测显示，27个样本中没有一个来自监测铜绿假单胞菌感染的2个特定无病原体（SPF）小鼠设施；未监测铜绿假单胞菌感染的SPF小鼠设施的12个样本中有1个样本和传统小鼠设施的7个样本中有2个样本铜绿假单胞菌阳性。相比之下，铜绿假单胞菌没有检测到任何样品通过传统的培养试验。因此，该比色LAMP法是一种简便、快速的铜绿假单胞菌检测方法。(C) 2010 Elsevier B.V.版权所有
A colorimetric loop-mediated isothermal amplification (LAMP) assay with hydroxy naphthol blue was designed to amplify a region in the outer membrane lipoprotein (oprL) gene of Pseudomonas aeruginosa. The LAMP assay showed 100% specificity for the serogroup and other bacteria, and the sensitivity was 10-fold higher than that of the PCR assays. The LAMP assay could detect P. aeruginosa inoculated in mouse feces at 130 colony-forming units (CFU)/0.1 g feces (3.25 CFU/reaction). The assay was completed within 2 h from DNA extraction. In a field trial, the LAMP assay revealed that none of the 27 samples was obtained from 2 specific pathogen-free (SPF) mouse facilities that were monitoring infection with P. aeruginosa; 1 out of 12 samples from an SPF mouse facility that was not monitoring infection with P. aeruginosa and 2 out of 7 samples from a conventional mouse facility were positive for P. aeruginosa. In contrast, P. aeruginosa was not detected in any of the samples by a conventional culture assay. Thus, this colorimetric LAMP assay is a simple and rapid method for P. aeruginosa detection. (C) 2010 Elsevier B.V. All rights reserved.